Plant biotechnology: Plant biotechnology: a key technology in the 21st centurya key technology in the 21st century

August, 2009

Atsuhiko ShinmyoNara Institute of Science and Technology

E- mail shinmyou@bs.naist.jp

ArgentineArgentine--Japan Work ShopJapan Work Shop

●

生駒市

◎

奈良市

Todaiji：Big Budda

Yakushiji

● Tokyo

Osaka ●

Kohukuji：Asura

Kyoto

Osaka

Nara

Tohin garden

Sujakumon

Reconstruction of Taikyokuden

Reconstruction of Kentoshisen

Nara is the 1Nara is the 1stst Capital in Japan established in AD 710Capital in Japan established in AD 710

2010:1300 years anniversary2010:1300 years anniversary

Nara Institute of Science and TechnologyNara Institute of Science and Technology

Information Science

Material science

Bioscience

Established in 1991Established in 1991

Prof. Prof. ShinsukeShinsuke Yamanaka of Yamanaka of iPSiPS was grown in NAIST.was grown in NAIST.

GDP (gross domestic product) Ranking

Country Population GDP Country Population* GDP**

2000 (M) 2006 (kB$) 2050 (M) 2050 (k B$)

1 USA 285 13.19 China 1,409 70.7

2 Japan 127 4.38 USA 402 38.5

3 Germany 82 2.89 India 1,658 37.7

4 China 1,270 2.67 Brazil 254 11.4

5 UK 59 2.37 Mexico 132 9.3

6 France 59 2.23 Russia 108 8.6

7 Italy 58 1.85 Indonesia 297 7.0

8 Canada 31 1.27 Japan 95 6.7

9 Spain 40 1.23 UK 68 5.1

10 Brazil 174 1.07 Germany 74 5.0

*Ministry of International Affairs and Communications, Japan **Goldman Sachs (2007)

Potential of underused renewable energy sources

Hydro Tides &currents

Wind Geo-thermal

Solar Currentuse

0.1

1

10

100

1000

10000

100000

1000000

TW

C. Somerville (NEDO Workshop, Osaka, 2006, 9, 14)TW (tera watt)=1,000 Billion watt

Plantbiomass

Use of Oil Products in Japan（2006）

Gasoline

Naphtha

Jet fuelKerosene

Diesel

Heavy oil

229 million kl

Ethanol

Annual Report of Resources and Energy

BioBio--dieseldiesel

Lignin

Industrial materials by

plants

Ethanol is an Excellent Transportation Fuel Ethanol is an Excellent Transportation Fuel Compared to Gasoline Compared to Gasoline

oo Has a higher octane rating; causes a disproportionate increase Has a higher octane rating; causes a disproportionate increase in octane rating when blended with gasoline; replaced in octane rating when blended with gasoline; replaced tetraethyl lead as octane enhancertetraethyl lead as octane enhancer

oo Burns with greater efficiency Burns with greater efficiency oo Produces lower amounts of ozone precursors, thus decreasing Produces lower amounts of ozone precursors, thus decreasing

air pollution, no air pollution, no SOxSOx and and NOxNOxoo Lower net C0Lower net C022 contribution to atmospherecontribution to atmosphereoo Free from sea water pollutionFree from sea water pollutionoo More favorable trade balanceMore favorable trade balanceoo Enhanced energy security Enhanced energy security oo Major new crop for depressed agricultural economy Major new crop for depressed agricultural economy

(Wyman and (Wyman and HinmanHinman, 1990;Lynd et al;1991; Greene et al., 2004), 1990;Lynd et al;1991; Greene et al., 2004)

Bioenergy Research Centerin US: $200 M （2007－2012）

• Support R&D projects for ethanol production for

automobile, production of fine chemicals and

industrial materials from biomass

• President Bush ： cost down of cellulose-ethanol to

that of gasoline in 2012

• Cut 20% of gasoline within 10 years

• Domestic supply of renewable clean energy

Bioenergy Science Center Great Lakes Bioenergy Research CenterJoint Bioenergy Institute

Possibility of replacement of gasoline to ethanolPossibility of replacement of gasoline to ethanol

World Japan

Gasoline consumption 2.6 B kl (100%) 60 M kl (100%)

Starch production 2.8 B ton 17 M tonEthanol production 1.8 B kl (70%) 11 M kl (18%)

Unused biomass 52 B ton 220 M tonEthanol production 21 Bkl (800%) 88 M kl (150%)

Waste biomass 4.3 B ton 49 M tonEthanol production 2.3 B kl (90%) 20 M kl (33%)

Unused biomass: wild forest, weeds, wastes

640 kl ethanol is produced from 1 ton starch.

400 kl ethanol is produced from 1 ton rice straw.

Y. Nabeshima: Metabolic Engineering of Plants (2002)

H H CH3O•CO-R1HC•O•CO-R1 HC•OHHC•O•CO-R2 + 3CH3OH → HC•OH + CH3O•CO-R2 HC•O•CO-R3 HC•OH

H H CH3O•CO-R3

Oil Methanol Glycerol Fatty acid methyl ester

(BioBio--diesel fuel)diesel fuel)

Enzymatic production with lipase will be better.

BioBio--diesel fueldiesel fuel

Law materials: plant and animal oilLaw materials: plant and animal oil

Catalyst in alkaline condition

Annual production of oil biomassAnnual production of oil biomass

Production (Production (MtonMton/Y)/Y) Oil (Oil (MtonMton)) Oil yield (ton/ha)Oil yield (ton/ha)

SoybeanSoybean 2.142.14 17.6 0.35 17.6 0.35

RapeseedRapeseed 0.46 0.46 12.012.0 0.640.64

Oil palmOil palm 0.55 23.0 0.55 23.0 4.94.9

SunflowerSunflower ーー 6.0 0.436.0 0.43

JatrophaJatropha ーー ーー 1.751.75

Attractive oil plant, Jatropha curcas

Origin: Central AmericaGrow in semi-drought, active growth over 20℃, 3～5m height, grow 50 yearsOil content in seed, 30～40％

Non-food, because of toxic compound,pholbol ester

Oil yield, 1.75 ton/ha/year, next of oil palm

Annual consumption of diesel oil in the world : 1.5 B kl

Jatropha oil production: 1.9 kl/ha (Density of bio-diesel : 0.93)

Cultivation land required : 800 MhaSemi-dry land in on the earth :

3,400 Mha

B747-300

Haneda, Tokyo

2009, 1, 30

Bio-flight :Test flight by bio-diesel fuel was succeeded.

The third engine of B747-300 was drived by pure bio-jet fuel (Camelinaoil:84％, Jatropha oil:15％, algae oil:1％ mixture).

2008, 2 2008, 12 2009, 1, 7

babasu oil and coconut oil 80% jet fuel 20%

Jatropha oil 50% jet fuel 50%

Jatropha oil algal oil 50% jet fuel 50%

Comparison of process of bio-fuel production

Most important factor is a cost of law materials.

starch 1.6 kg → ethanol 1lstarch 25 yen/kg → ethanol 40 yen/l

Fat

fatty acid methyl ester

esterification(bio-diesel)

（heavy oil A）(direct)

No energy inputNo energy input

ethanolabsolute ethanol(gasoline)

fermentationconcent-ration

high energy inputhigh energy inputinhibitorinhibitor

Sucrose

Starch

Cellulosicbiomass

amylase

cellulase, hemi-cellulase

pre-treatment

glucose

glucosexylose

direct

Soil: N, P, K, S, Me, H2O

Atmosphere

CO2

Starch, cellulose

(C6H12O6)n

Fatty acidCH3(CH2)nCOOH

CO2

H2O

Chemical energy

Solar energy

O2

Other components

Return to soil

O2

Recycle system utilizing plant biomass energy

Plant biomass

Sustainable world！

12011010090 80706050403020100

Present use

Totalplant

biomassUsed biomass (7%)(food, feed, wood, pulp, textile)

Required for maintenance of forest (33%)

Unused biomass (60%)forestry: agriculture: stock raising

24 : 41 : 35

Increase of biomass (12%)

Ene

rgy

(TW

)Plant Biomass EnergyPlant Biomass Energy

Recombinant DNA Recombinant DNA technologytechnology

Useful genes

(any gene from any organism)

×Breeding by Breeding by

crossingcrossingwithin close relatives

Accidental result

Messiah of humans!Messiah of humans! Recombinant DNA technologyRecombinant DNA technology

Drought

Salt

Temperature

Active oxygen

Acid rain

Disease

Insects

Poor nutrition

Stress to plant

病気

害虫

雑草

干ばつ土壌悪化

冷害

水害 収穫

その他

Yield

DiseaseInsect

Weed

DroughtSoil deterioration

Cold weather

Flood

Others

Decrease of productivity

of plant by stress in US

Boyer:Science 1982

Eucalyptus

50% of pulp materials

Growth: 5 m/year

Growth of Eucalyptus in acidic soil by citrate secretion

Utilization of

rock phosphate

Ohji Paper Co.

Leaf

110%

Root

124%

Wild Transgenic

Wild type tobacco

0 2 00 4 00 6 00 8 00 1,000 1,200 1,400 1,600

Transformant

-2

0

2

4

6

8

10

12

14

***

**

*

**

***

16

18

20

22

**

** *

*

*

*

*

**

Light intensity (µmol photons m-2 s-1)

Phot

osy n

thet

i c a

c tiv

it y （

µmo l

CO

2 m

-2s-1

) ）

FBP/SBPase gene Activation of RuBisCO

Wild type

Increase of photosynthesis by chloroplast transformationYokota, NAIST（Jap. Pat. App. 2004-59513）

Transformant

Gene for synthesize flowering hormone, Gene for synthesize flowering hormone, florigeneflorigene

0

10

20

30

40

50

60

70

Wild Hd3a

Day

for f

low

erin

g (d

ay) Shortening of flowering time in rice

Shortening of harvesting time to 60%→ Rice production : 3 times

in Japan per year→ Extend to wheat, corn, soybean, so on

Shimamoto andTamaki, NAIST (2007)

Water DryWild water melon

The gene Wild

Trans-genic

Arabidopsis

Gene for extension of root Gene for extension of root from water melon in from water melon in

BotswanaBotswana desertdesert

Yokota and Akashi, NAIST （2007）

Strategy for increase of biomass production

1) Increase of cultivation landUtilization of dry and salty land, high/low temperature area/period, and acidic/alkaline soil

2) Increase of productivity per unit landIncrease of photosynthesis, CO2 fixation, growth rate,

and size of seeds/tuber

Shortening of harvesting period

3) Molecular breedingStress-resistance and increase of productivity

Total land on the earth : 12.8 billion hectare(except lake, river and pond)

Agricultural land

Forest

DesertDry land

Urban

Frozen land

Mountain Others 1.525 B ha

3.43 B ha

Acidic soil

(42% of agricultural land)

Alkaline soil

Salty soil

Poor land

Increase of plant productivity by rDNA technology

Future

Now

rDNA technology

Geneticalmaximum

Stress

Productivity

Stress-resistantgenes

Metabolicgenes

Productivity

Stress

Productivity

Stress

2. Technology for regulation of biosynthesis in useful plantsTechnology : genes for metabolism, analysis of metabolites,

identification of key gene, transformation of useful plants, increase of productivity, cultivation

Materials : Eucalyptus, licorice, rubber tree, Eucommia, flax

Product : pulp, rubber, terpenoid, steroid, carotenoid, hyaluronic acid

1. Analysis of biosynthesis of metabolites in model plantsTechnology : Provide basic resources for biosynthetic process

cDNA, gene expression, gene function, regulation of gene expression, microarray, metabolome, data base

Materials : Arabidopsis thaliana, Lotus comiculatus

METI-NEDO Plant Project （2002～2009）

GPP

FPP

trans-polyisoprenen

OPP

cis-polyisoprene

OPP

OPP

OPP

n

H

OPP

OPP

IPP isomerase

IPP

DMAPP

poly

pren

yl-P

P sy

ntha

se

IPP

Rubber( polyisoprenoid) biosynthetic pathway

IPP

(C5)(C5)

(C10)

(C15)

IPP

Rubber tree

Bridgestone

Tochu (Eucommia ulmoides)

Hitachi

From Bioscience to Biotechnology in PlantsFrom Bioscience to Biotechnology in PlantsMany important genes have been isolated from model plants, such as Arabidopsis etc., and analyzed their functions. (stress-resistance, growth stimulation, biosynthesis of metabolites, transcription factors, regulatory elements)

• Application to useful plants

• Genetically modified plants by multi-genes

• Quantitative regulation of gene expression

(Bioscience)

(Biotechnology)

Basic and applied life scienceBasic and applied life science

Animal scienceAnimal science Plant sciencePlant science

AppliedApplied

BasicBasic

Human

Model animals (various)

Various plants

Model plants(Arabidopsis)

Black GoldBlack Gold

Green GoldGreen Gold

20 C20 C

21 C21 C

Blue Gold

OilOil--producing producing

countries became rich.countries became rich.

Countries abundant Countries abundant

biomass and strong to biomass and strong to

biotechnologybiotechnology will be will be

happy.happy.

PetroleumPetroleum

BiomassBiomass